This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. To advance general biomedical training we continue to field questions from researchers from all over world on FRET analysis and general questions on our imaging protocols. We have developed an online video that describes our techniques in detail. We have submitted the code for our software, accompanied with a detailed user manual, to the Matlab Central website for distribution with a GNU general public license.